A covalent link between the chromophore and the protein backbone of bacteriorhodopsin is not required for forming a photochemically active pigment analogous to the wild type

Biochemistry. 1994 Mar 1;33(8):1971-6. doi: 10.1021/bi00174a001.

Abstract

Bacteriorhodopsin pigments lacking the retinal-Lys-216 covalent bond were prepared by reconstituting the K216G mutant protein with retinal alkylamine Schiff bases. The procedure follows the approach of Zhukovsky et al. [Zhukovsky, E., Robinson, P., & Oprian, D. (1991) Science 251, 558-560] in the case of visual (rhodopsin) pigments. Reconstitution leads to a mixture of three pigments. One of them, bR(K216G)/566a, absorbs (pH = 6.9) at 566 nm. Its absorption is pH-dependent, exhibiting a purple to blue transition. The pigment's laser-induced photocycle patterns are similar to those of wild-type all-trans-bR. A second component, bR(K216G)/566b, exhibits an independent photocycle reminiscent of that of wild-type 13-cis-bR. A third pigment component, bR(K216G)/630, absorbs around 630 nm. Experiments in the presence of a pH dye indicator show that illumination of bR(K216G)/566 produces a detectable proton gradient. It is concluded that a covalent bond between the retinal chromophore and the protein backbone is not a prerequisite for the basic structure and photochemical features of bR or for its proton pump activity.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacteriorhodopsins / chemistry*
  • Halobacterium salinarum / chemistry
  • Photochemistry
  • Pigments, Biological / chemistry*
  • Proton Pumps
  • Retinaldehyde / chemistry
  • Schiff Bases / chemistry

Substances

  • Pigments, Biological
  • Proton Pumps
  • Schiff Bases
  • Bacteriorhodopsins
  • Retinaldehyde